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Sensors 2015, 15(2), 3896-3910; doi:10.3390/s150203896

Nano-Magnetic Immunosensor Based on Staphylococcus Protein A and the Amplification Effect of HRP-Conjugated Phage Antibody

Research Institute of Chemical Defence, State Key Laboratory of NBC Protection for Civilian, Beijing 102205, China
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Author to whom correspondence should be addressed.
Received: 26 November 2014 / Accepted: 2 February 2015 / Published: 9 February 2015
(This article belongs to the Special Issue Immunosensors 2014)
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Abstract

In this research, super-paramagnetic Fe3O4 nanoparticles (magnetic particles) were coated with Staphylococcus protein A (SPA) and coupled with polyclonal antibody (pcAb) to construct magnetic capturing probes, and HRP-conjugated phage antibody was then used as specific detecting probe to design a labeled immunosensor for trace detection of Staphylococcus aureus enterotoxin B (SEB). The linear detection range of the sensor was 0.008~125 µg/L, the regression equation was Y = 0.487X + 1.2 (R = 0.996, N = 15, p < 0.0001), the limit of detection (LOD) was 0.008 µg/L, and the limit of quantification (LOQ) was 0.008 µg/L. HRP-conjugated phage antibody, SPA and magnetic particles can enhance the sensitivity 4-fold, 3-fold and 2.6-fold higher, respectively. Compared with conventional double-antibody sandwich ELISA, the detection sensitivity of the sensor was 31-fold higher resulting from the integrated amplifying effect. The immunosensor integrates the unique advantages of SPA-oriented antibody as magnetic capturing probe, HRP-conjugated phage antibody as detecting probe, magnetic separation immunoassay technique, and several other advanced techniques, so it achieves high sensitivity, specificity and interference-resistance. It is proven to be well suited for analysis of trace SEB in various environmental samples with high recovery rate and reproducibility. View Full-Text
Keywords: Staphylococcus protein A; HRP-conjugated phage antibody; signal amplification; nanomagnetic immunosensor; Staphylococcus aureus enterotoxin B Staphylococcus protein A; HRP-conjugated phage antibody; signal amplification; nanomagnetic immunosensor; Staphylococcus aureus enterotoxin B
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Mu, X.; Tong, Z.; Huang, Q.; Liu, B.; Liu, Z.; Hao, L.; Zhang, J.; Gao, C.; Wang, F. Nano-Magnetic Immunosensor Based on Staphylococcus Protein A and the Amplification Effect of HRP-Conjugated Phage Antibody. Sensors 2015, 15, 3896-3910.

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